The prior art has developed a wide variety of test means for the determination of specific constituents in liquids such as urine and blood. These have taken a variety of forms, one of the most popular being reagent impregnated test strips of the dip-and-read type, certain of which are useful for the determination of such constituents as glucose, protein, occult blood, and the like in body fluids, whereas others are useful for the determination of various constituents in other liquids, such as swimming pool water, cutting fluid, and the like.
Such prior art test systems have conventionally been of the type which include in the reagent composition one or more chromogenic redox indicators which are either directly responsive to the analyte to be determined or are combined with and react to the product of an analyte responsive system. Recently, methods have been developed whereby chemiluminescent techniques have been used for determination of glucose in blood [Bostik D. T. et al. Anal. Chem., 47:447-452 (1975)] and in urine [Williams, D. C. et al. Clin. Chem., 22:372-374 (1976)]. These determinations have made use of glucose oxidase immobilized to a column through which a test sample is passed by positive pressure using an infusion pump and syringe. As the glucose sample enters the column, hydrogen peroxide is generated and carried out of the column with the column effluent to an optically clear cell in which it reacts with luminol ferricyanide in a liquid system. The chemiluminescence produced is detected by a separate photomultiplier tube which is attached to the face of the cell. The signal is then amplified by various photometric preamplifiers and recorded by a potentiometric recorder.
More recently an automated chemiluminescent method for detemining nicotinamide adenine dinucleotide, such as is used in lactate dehydrogenase determinations, was published by Williams, D. C. et al, Anal. Chem., 48: 1478-1481 (1976). A segmented flow system driven by a peristaltic pump was used.
Another apparatus which has been suggested for chemiluminescent determination is simply prepared by injecting a sample and chemiluminescent reagents into a sealed container surrounded by photographic film and measuring the film exposure as a function of concentration. [Seitz, W. R. et al. Anal. Chem., 46:188-202, at 191-192 (1974)].
Coffman, U.S. Pat. No. 3,239,406 discloses a chemiluminescent tape useful as a marker. Upon exposure to air the tape chemiluminesces for different periods of time and at different levels of illumination depending upon the type and amount of chemiluminescent composition incorporated in the structure. The tape comprises at least one layer or surface which is adhesive to other surfaces and which has at least a surface impregnated with a chemiluminescent composition containing at least one peraminoethylene and a strippable film overcoat or removable envelope to protect the peraminoethylene composition from exposure to oxygen prior to use.
Cavanagh, U.S. Pat. No. 3,923,462, discloses an automated apparatus for the detection of ozone in ambient air. A sample of air is passed through a light tight enclosure where it reacts with a material such as Rhodamine B, which luminesces in the presence of ozone, or a material which normally luminesces (such as in black light) and is quenched in the presence of ozone. Photographic film is positioned in the enclosure and spaced apart from the chemiluminescent system. The film is in exposed relationship to the luminescent reaction inside of the light tight enclosure. The pressure, such as atmopheric pressure, of the substance to be detected must be determined independently of film density, thus requiring two separate measurements and use of sophisticated and expensive equipment.
Thus, it can be seen that the application of chemiluminescence has been as markers, indicators of gas content and, in expensive continuous flow column techniques, luminescent reactions have been used in analytical chemistry. Despite the development of the dip-and-read test device industry and the attempts at application of luminescence reactions to analytical chemistry it is evident that methodologies to which each are applicable have been limited in scope. Substantial areas of analysis have not heretofore been possible with conventional dip-and-read test devices because of the detection ranges to which chromogenic indicators are limited. Likewise the areas of analysis to which prior art luminescent systems have been applicable is limited by the size, expense, complexity and susceptibility to interference which are characteristic of the methods disclosed.